Main description:

This detailed book explores the fundamentals of myofibroblast biology in tissue repair, fibrosis, and tumors as well as providing cutting-edge laboratory methods used to investigate myofibroblast functions in physiological and pathological settings in vitro and in vivo, as written by leading academic scientists. Section I of this volume focuses on fundamental methods to study myofibroblast biology and covers topics such as methods for detecting myofibroblasts and senescent myofibroblasts in cell culture and histology, single cell RNA sequencing to identify myofibroblast subsets in fibrotic tissues, and functional assays to assess TGF- activation, myofibroblast apoptosis, or matrix deposition and crosslinking. Section II discusses methods to investigate the mechanobiology of myofibroblasts in vitro, including the fabrication of functional hydrogels with tunable stiffness, the use of atomic force microscopy to characterize matrix and cellular stiffness, as well as molecular assays to assess fibroblast mechanotransduction pathways and durotaxis. Section III describes multiple animal models to investigate myofibroblast functions across organs in vivo as well as human organoid systems, precision tissue slices and decellularized 3D tissue scaffolds to assess myofibroblast functions in relevant human ex vivo models. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and extensive, Myofibroblasts: Methods and Protocols is an essential collection for researchers delving into the processes and effects of these important cells.

Contents:

1. 50 Years of Myofibroblasts: How the Myofibroblast Concept Evolved

Giulio Gabbiani

Section I: Fundamental Methods to Study Myofibroblast Biology

2. Myofibroblast Functions in Tissue Repair and Fibrosis: An Introduction

Victor J. Thannickal

3. Myofibroblast Markers and Microscopy Detection Methods in Cell Culture and Histology

Fereshteh Younesi, Dong Ok Son, Joao Firmino, and Boris Hinz

4. Fibroblast and Myofibroblast Subtypes: Single Cell Sequencing

Hesham Soliman, Lin Wei Tung, and Fabio M.V. Rossi

5. Myofibroblast Adhesome Analysis by Mass Spectrometry

Christopher A. McCulloch

6. Myofibroblast TGF- Activation Measurement In Vitro

Joanne Porte, Gisli Jenkins, and Amanda L. Tatler

7. Contraction Measurements Using Three-Dimensional Fibrillar Collagen Gel Lattices

James Griffith and William L. Berry

8. Techniques to Assess Collagen Synthesis, Deposition, and Cross-Linking In Vitro

Tamara Rosell-Garcia and Fernando Rodriguez-Pascual

9. Methods for Studying Myofibroblast Apoptotic Pathways

Yan Zhou and David Lagares

10. Determination of Senescent Myofibroblasts in Precision-Cut Lung Slices

Tamara Cruz, Ana L. Mora, and Mauricio Rojas

11. The Scar-in-a-Jar: In Vitro Fibrosis Model for Anti-Fibrotic Drug Testing

Simon Stebler and Michael Raghunath

Section II: Myofibroblast Mechanobiology

12. Why Stress Matters: An Introduction

Daniel J. Tschumperlin

13. Soft Substrate Culture to Mechanically Control Cardiac Myofibroblast Activation

Natalie M. Landry, Sunil G. Rattan, and Ian M.C. Dixon

14. Quantitative Analysis of Myofibroblast Contraction by Traction Force Microscopy

Shuying Yang, Fernando R. Valencia, Benedikt Sabass, and Sergey V. Plotnikov

15. Nucleocytoplasmic Shuttling of the Mechanosensitive Transcription Factors MRTF and YAP/TAZ

Michael Kofler and Andras Kapus

16. Atomic Force Microscopy for Live-Cell and Hydrogel Measurement

Alexander Whitehead, Natalie J. Kirkland, and Adam J. Engler

17. Method for Investigating Fibroblast Durotaxis

Hossam Kadry, David Lagares, and Taslim A. Al-Hilal

18. Decellularized Extracellular Matrix (ECM) as a Model to Study Fibrotic ECM Mechanobiology

Chiuan-Ren Yeh, Grace C. Bingham, Jagathpala Shetty, Ping Hu, and Thomas H. Barker

19. Fibrosis on a Chip for Screening of Anti-Fibrosis Drugs

Mohammadnabi Asmani and Ruogang Zhao

Section III: Animal and Human Models to Study Myofibroblast Biology

20. Animal and Human Models of Tissue Repair and Fibrosis: An Introduction

David Lagares and Boris Hinz

21. Mouse Models of Lung Fibrosis

Olivia Mekhael, Safaa Naiel, Megan Vierhout, Aaron Hayat, Spencer Revill, Soumeya Abed, Mark Inman, Martin Kolb, and Kjetil Ask

22. Mouse Models of Kidney Fibrosis

Rafael Kramann and Sylvia Menzel

23. Mouse Models of Liver Fibrosis

Aashreya Ravichandra and Robert F. Schwabe

24. Mouse Models of Muscle Fibrosis

Antonio L. Serrano and Pura Munoz-Canoves

25. Mouse Models of Skin Fibrosis

Aleix Rius Rigau, Markus Luber, and Joerg H.W. Distler

26. Mouse Models of Intestinal Fibrosis

Jiannan Li, Dina Dejanovic, Megan T. Zangara, Jyotsna Chandra, Christine McDonald, and Florian Rieder

27. A Rodent Model of Hypertrophic Scarring: Splinting of Rat Wounds

Dong Ok Son and Boris Hinz

28. Three-Dimensional Model of Hypertrophic Scar Using a Tissue-Engineering Approach

Veronique J. Moulin

29. Methods for the Study of Renal Fibrosis in Human Pluripotent Stem Cell-Derived Kidney Organoids

Alejandro Moran-Horowich and Dario R. Lemos

30. Decellularized Human Lung Scaffolds as Complex Three-Dimensional Tissue Culture Models to Study Functional Behavior of Fibroblasts

Gerald Burgstaller, Michael Gerckens, Oliver Eickelberg, and Melanie Koenigshoff